细菌表达重组朊病毒蛋白的转化。
Conversion of bacterially expressed recombinant prion protein.
机构信息
Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
出版信息
Methods. 2011 Mar;53(3):208-13. doi: 10.1016/j.ymeth.2010.12.013. Epub 2010 Dec 19.
Abstract
The infectivity associated with prion disease sets it apart from a large group of late-onset neurodegenerative disorders that shares the characteristics of protein aggregation and neurodegeneration. The unconventional infectious agent, PrP(Sc), is an aberrantly folded form of the normal prion protein (PrP(C)) and the PrP(C)-to-PrP(Sc) conversion is a critical pathogenic step in prion disease. Using the Protein Misfolding Cyclic Amplification technique, we converted folded bacterially expressed recombinant PrP into a proteinase K-resistant and aggregated conformation (rPrP-res) in the presence of anionic lipid and RNA molecules. Moreover, high prion infectivity was demonstrated by intracerebral inoculation of rPrP-res into wild-type mice, which caused prion disease with a short incubation period. The establishment of the in vitro recombinant PrP conversion assay makes it feasible for us to explore the molecular basis behind the intriguing properties associated with prion infectivity.
摘要
朊病毒病的感染性使其有别于一大类具有蛋白聚集和神经退行性变特征的晚发性神经退行性疾病。非常规感染因子 PrP(Sc)是正常朊病毒蛋白 (PrP(C))的异常折叠形式，而 PrP(C)到 PrP(Sc)的转化是朊病毒病的关键致病步骤。使用蛋白质错误折叠循环扩增技术，我们在阴离子脂质和 RNA 分子的存在下，将折叠的细菌表达重组 PrP 转化为蛋白水解酶抗性和聚集构象（rPrP-res）。此外，通过将 rPrP-res 脑内接种到野生型小鼠中，证明了高朊病毒感染力，导致潜伏期短的朊病毒病。体外重组 PrP 转化测定法的建立使我们能够探索与朊病毒感染力相关的有趣特性的分子基础。
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